川芎高频再生体系的建立

为建立川芎(Ligusticum chuanxiong Hort.)高频再生体系,优化了诱导和分化培养基及培养条件。以叶柄为外植体,以MS为基本培养基,KT 2.0 mg/L+IAA 0.5 mg/L的激素组合对不定芽分化最有利。在此基础上,针对外植体来源、培养条件和愈伤组织继代时间3个因素进行优化。结果表明:采用川芎无菌苗叶柄作为外植体,黑暗条件下诱导出愈伤组织,再在光照下继代培养15 d后转入分化培养基中对不定芽诱导最为有利,分化率为44.4%。分化后得到的不定芽在含NAA 0.5 mg/L和IBA 0.5 mg/L的 1/2MS培养基上生根率达90%,移栽存活率为95%。     

长柄双花木愈伤组织诱导及植株再生

以长柄双花木当年生嫩梢上的叶柄、嫩茎、嫩叶为外植体,对影响长柄双花木愈伤组织诱导和继代、分化主要因素进行研究。结果表明:在培养基MS+NAA0.5mg/L+2,4-D2.0mg/L上,三种外植体均可诱导出愈伤组织,其中叶片愈伤组织诱导率最高。该培养基还可作为愈伤组织继代培养基,但继代培养周期不超过2周。愈伤组织接种在MS+BA2mg/L上分化不定芽,根的诱导在1/2MS+IBA0.5mg/L培养基上进行。     

玉竹的组织培养与快速繁殖

以玉竹[Polygonatum odoratum (Mill.) Druce]根状茎、叶片和茎段为外植体,于附加不同激素配比的MS培养基中诱导愈伤组织、不定芽和不定根,探讨增殖培养和植株再生的条件.结果表明,叶片和茎段外植体诱导愈伤组织和芽的分化率很低;而根状茎外植体易于培养,有较高的诱导率和增殖倍数,其愈伤组织、不定芽和不定根的诱导率分别可达87%、90%和99%以上.适宜根状茎外植体愈伤组织诱导的培养基为MS+1.0 mg/L 6-BA+0.5 mg/L NAA,有利于增殖和丛生芽分化的培养基为MS+2.0 mg/L 6-BA+0.5 mg/L IBA和MS+3.0 mg/L 6-BA+0.1 mg/L NAA,而1/2MS+3.0～5.0 mg/L NAA适宜诱导试管苗生根培养.试管苗的移栽成活率可达85%以上.     

黄花补血草愈伤组织的诱导和植株再生

以黄花补血草(Limonium aureum(L.)Hill.)无菌苗为材料,研究了不同激素配比条件下不同外植体愈伤组织的诱导及植株再生.结果表明,黄花补血草无菌苗叶片、叶柄和幼根均可作为离体培养的外植体,但叶片和叶柄的诱导率明显高于幼根.将外植体接种于分别添加0.5 mg/L NAA或1.0 mg/L 2,4-D或0.3-0.5 mg/L 6-BA 0.5-2.0 mg/L NAA的MS培养基上,经过14-28 d培养后,可脱分化产生乳白色、红色或浅绿色颗粒状或致密愈伤组织,频率达到70%以上.在MS 0.3 mg/L 6-BA 1.0 mg/L NAA培养基上,红色和绿色颗粒状愈伤组织经过1-2次继代后,均可分化产生不定芽,进而形成丛生芽,分化率达到100%.将高约3 cm的丛生芽切下,接种于分别添加0.5 mg/L IAA或0.5 mg/L IBA的1/2 MS培养基上可产生不定根,获得完整的再生植株.     

梨蒴珠藓愈伤组织诱导和配子体再生研究

以梨蒴珠藓无菌藓株为外植体诱导愈伤组织和配子体再生,接种于含不同激素组合的MS和Knop固体培养基上,分别进行愈伤组织和不定芽的分化,并探讨愈伤组织诱导和配子体再生的适宜培养条件.结果显示,愈伤组织诱导的最佳培养基是MS+0.5 mg/L BA+0.1 mg/L 2,4-D,愈伤组织诱导率为33.3%;不定芽诱导的最佳...     

非洲菊试管苗叶柄愈伤组织的诱导与分化研究

以非洲菊(Gerbera janesonii Bolus)品种‘Sunanda'试管苗幼叶的带叶叶柄为材料,研究了培养基、外植体类型和继代次数等因素对愈伤组织诱导及分化的影响.结果表明:培养基上附加不同植物生长调节剂所诱导出的愈伤组织在形态和分化能力上存在显著差异,叶柄基部诱导愈伤组织的最适培养基是MS+TDZ 0.3 mg L-1+NAA0.1 mg L-1.外植体以叶片长度＞0.5 cm,叶片已舒展,颜色嫩绿的幼叶最佳,继代培养基以MS+KT 1.0 mg L-1较适宜,愈伤组织在分化培养基MS+6-BA 2.0 mg L-1+NAA 0.1 mg L-1上分化出芽的同时还会增殖,分化率达87.4%,继代培养2次的愈伤组织分化率可提高至95%.不定芽在生根培养基1/2MS+IBA0.6 mg L-1上的生根率达100%,试管苗移栽45 d后,成活率达97%以上.     

菘蓝两种外植体愈伤组织诱导研究

为寻找菘蓝愈伤组织诱导的最佳外植体,最佳培养基及愈伤组织继代增殖的最佳培养基,选用叶片和叶柄两种外植体,在添加不同种类植物激素处理组合的培养基上,对不同外植体进行愈伤组织的诱导和继代增殖研究.结果表明叶片比叶柄愈伤组织的诱导能力强,表现为出愈时间早,诱导率高,出愈多且质量好.叶片愈伤组织诱导培养基:MS 6-BA1.0mg/L 2,4-D0.3～0.5mg/L;继代增殖培养基:MS 6-BA0.5mg/L 2,4-D0.3mg/L.因此,采用菘蓝叶片为外植体,能高效的诱导出愈伤组织并能快速增殖.     

樱桃砧木Colt离体叶片再生

以樱桃砧木Colt试管苗的叶片为外植体,通过先诱导愈伤组织分不定芽以及叶片直接分化不定芽两种途径诱导再生,结果表明,在MS附NAA1．0mg/L、KT3．0mg/L、ZT0．25mg/L培养基中,愈伤诱导率可达100％;诱导的愈伤MS附加NAA0．2mg/L、IAA0．5mg/L、6－BA0．5mg/L、KT1．0mg/L,GA0．5mg/L培养基中,不定芽分化率为21．3％,在MS附加6-BA6．0mg/L、NAA1．0mg/L、GA0．5mg/L中,叶片一叶柄不定芽诱导率可达48．3％。     

樱桃砧木Colt离体叶片再生

以樱桃砧木Colt试管苗的叶片为外植体 ,通过先诱导愈伤组织分化不定芽以及叶片直接分化不定芽两种途径诱导再生。结果表明 :在MS附加NAA 1 0mg/L、KT3 0mg/L、ZT0 2 5mg/L培养基中 ,愈伤诱导率可达 1 0 0 % ;诱导的愈伤在MS附加NAA 0 2mg/L、IAA0 5mg/L、6 BA 0 5mg/L、KT 1 0mg/L、GA 0 5mg/L培养基中 ,不定芽分化率为 2 1 3% ;在MS附加 6 BA 6 0mg/L、NAA 1 0mg/L、GA 0 5mg/L中 ,叶片 -叶柄不定芽诱导率可达 48 3%。     

万寿菊杂交亲本的离体培养

以万寿菊‘钻石’雄性不育株的幼嫩叶片和子房为外植体进行离体快繁,结果表明:培养基MS+0.8 mg·L-1 IAA+0.6 mg·L-1 6-BA+30 g·L-1蔗糖既有利于叶片愈伤组织的诱导也有利于不定芽的分化,愈伤组织诱导率达97.9％,不定芽诱导率达45.8％;培养基MS+0.5 mg·L12,4-D+0.5 mg·L-16-BA+ 30 g·L-1蔗糖为诱导子房愈伤组织的最佳培养基,出愈率为77.8％;培养基MS+0.05 mg·L -1 NAA+0.5 mg·L-1 6-BA+ 30g·L-1蔗糖为诱导子房愈伤组织不定芽分化的最佳培养基;将不定芽接至MS培养基上,7d后即可生出不定根,生根率可达98％,移栽成活率达90％以上.     

杜仲叶片和叶柄愈伤组织的诱导和植株再生

本实验以5~6年生杜仲叶片及叶柄为外植体,研究了杜仲愈伤组织诱导及植株再生的方法。结果表明:接种于补加NAA(2.0~4.0 mg/L)或BA(1.0 mg/L)+NAA(2.0~4.0mg/L)的MS培养基上的叶片和叶柄,经21~28d培养后,脱分化形成绿色或浅绿色致密愈伤组织,频率达到70%以上。绿色致密愈伤组织在补加BA(2.25~2.75 mg/L)+NAA(0.15 mg/L)的MS培养基上经过1~2次继代之后,即出现茎芽分化,频率在15%以上,只是其中许多都是畸形苗,正常苗频率较低。此问题尚在研究之中。选择生长健壮的再生植株,切除其基部愈伤组织,然后将切口浸泡在250mg/L无菌ABT生根粉溶液中3~5sec,再插入1/4强度无激素MS培养基中, 2~3周后,在苗基部长出1~3条白色粗壮的不定根,生根频率在60%以上。     

李砧木Marianna离体叶片再生体系优化研究

以李砧木‘Marianna’试管苗新梢顶端第1片叶为外植体,研究激素组合、基本培养基种类及外植体类型等对不定芽再生的影响。结果表明:1/2 MS基本培养基和WPM培养基再生率显著高于MS和SH培养基;叶片附带叶柄的外植体再生率和再生不定芽数显著高于叶柄和切除叶柄的叶片外植体;最佳再生培养基为1/2MS+2.0mg/L TDZ+0.1 mg/L IBA+0.25%琼脂+3.0%蔗糖,最高再生率和再生不定芽数分别为81.7%和7.46±1.38个;最佳生根培养基为1/2MS+0.5~1.0 mg/L IBA,能获得96.7%生根率、较高的生根数和根长。     

猕猴桃高频直接再生体系的建立

为了建立猕猴桃高频再生体系,以MS为基本培养基,猕猴桃(Actinidia deliciosaQinmei)茎及叶片为外植体,研究了2,4-D、6-BA和NAA在美味猕猴桃愈伤组织形成及分化过程中的作用。方差分析结果表明,6-BA能够显著促进愈伤组织形成,6-BA和NAA可以显著促进愈伤组织形成和分化,而2,4-D抑制愈伤组织形成。附加2.0 mg/L 6-BA、1.0 mg/L NAA和600 mg/L水解酪蛋白的MS培养基是茎段培养的最佳培养基,在该培养基上,以再生的无菌苗为起始材料,一个月时叶圆盘的直接再生频率达到100%,平均每个叶圆盘产生9.33个芽,其中23.21%芽高度超过0.5 cm。     

亚麻遗传转化体系的建立及几丁质酶基因导入的研究

报道了亚麻遗传转化体系的建立和几丁质酶基因对亚麻遗传转化的研究。亚麻下胚轴切段培养在不同激素浓度的ＭＳ培养基上,诱导分化出不定芽。最佳的激素组合是ＭＳ＋ＢＡ１ｍｇ／Ｌ＋ＩＡＡ０．５ｍｇ／Ｌ,分化频率可达９７％。亚麻的下胚轴经带有几丁质 根癌农杆菌感染后,在含有１００ｍｇ／Ｌ卡那霉素的选择分化培养基上,１４￣２１ｄ就能产生抗生小芽,小芽进一步伸长后可在１００ｍｇ／Ｌ卡那霉素的ＭＳ选择生根培养基（ＭＳ     

药用植物栝楼的组织培养及其表达蛋白的分析

对栝楼的快速繁殖、愈伤组织的诱导与再分化,以及不同培养体系中天花粉蛋白的表达进行了初步研究。结果表明：栝楼茎切段的腋芽和顶芽在MS+0.5、1.0mg/L 6-BA培养基上可以快速繁殖;组织培养苗的叶片切块在MS+4.0mg/L 6-BA +0.2mg/L IAA的培养基上可形成愈伤组织,该愈伤组织在30d后再分化为绿苗,绿苗分化率为0.25苗/外植体;绿苗转移至MS+0.1mg/L NAA的培养基可100%生根;生根苗移栽至土壤中100%成活;移栽成活的栝楼在30d后长出小块根,并检测到天花粉蛋白的表达。     

Influence of growth regulators on plant regeneration from epicotyl and hypocotyl cultures of two groundnut (Arachis hypogaea L.) cultivars

This study was designed to evaluate the effect of phytohormones on plant regeneration from epicotyl and hypocotyl explants of two groundnut (Arachis hypogaea) cultivars. Explants cultured on media with auxins and in combination with cytokinin produced high frequency of callus. After four weeks, callus from these cultures was transferred to medium with cytokinin and reduced auxin, shoot buds regenerated from the cultures. A high rate of shoot bud regeneration was observed on medium supplemented with 2.0 mg/L BAP and 0.5 mg/L NAA. Among the different auxins tested, NAA was found to be most effective, producing the highest frequency of shoot buds per responding cultures. Of the two explants tested, epicotyl was found to be best for high frequency shoot bud regeneration. Multiple shoots arose on MS medium supplemented with BAP or kinetin (1.0–5.0 mg/L) plus IBA (1.0 mg/L), with maximum production occurring at 5.0 mg/L. The elongated shoots developed rootsin vitro upon transfer to MS medium supplemented with NAA or IBA (0.5–2.0 mg/L) and kinetin (0.5 mg/L) for 15 days.In vitro produced plantlets, were transferred to soil and placed in a glasshouse developed successfully, matured, and set seeds.     

Organogenesis and Plantlet Regeneration in Vitro of Populus euphratica

The research of organogenesis and in vitro plantlet regeneration of Populus euphratica Oliver was carried out using the tender shoots from mature tree as initial explants and MS medium as the basic medium. The effects of plant growth regulators (PGR) on the regeneration were compared. The results showed that the concentration of PGR was not strictly required for the organogenesis of the excised organs and callus, but the ratio of BA to NAA was important. Calli could be induced from the excised leaves and stems cultured on the medium with 0.5 mg/L BA and 0.5 mg/L NAA. The embryonic callus could be multiplied in dark on the medium supplemented with 0.25 mg/L BA and 0.5 mg/L NAA. For the adventitious bud regeneration of the leaf and callus, supplement with 0.5 mg/L BA and 0.1 mg/L NAA was appropriate, giving a regeneration frequency of 82.9% and 100%, respectively. The suitable level of BA and NAA for the excised stem's was 0.1 mg/L and 0.01 mg/L respectively, yielding a regeneration frequency of 83 %. Rooting occurred on the MS medium with half strength of macronutrient and addition of 0.015 mg/L NAA, and the rooting rate could reach up to 86.2%. The techniques of somatic cell cloning of P. euphratica was established in vitro. The problems of deterioration of the subcultured shoots were also discussed.     

Efficient regeneration of sorghum, Sorghum bicolor (L.) Moench, from shoot-tip explant

Novel protocols for production of multiple shoot-tip clumps and somatic embryos of Sorghum bicolor (L.) Moench were developed with long-term goal of crop improvement through genetic transformation. Multiple shoot-tip clumps were developed in vitro from shoot-tip explant of one-week old seedling, cultured on MS medium containing only BA (0.5, 1 or 2 mg/l) or both BA (1 or 2 mg/l) and 2,4-D (0.5 mg/l) with bi-weekly subculture. Somatic embryos were directly produced on the enlarged dome shaped growing structures that developed from the shoot-tips of one-week old seedling explants (without any callus formation) when cultured on MS medium supplemented with both 2,4-D (0.5 mg/l) and BA (0.5 mg/l). However, the supplementation of MS medium with only 2,4-D (0.5 mg/l) induced compact callus without any plantlet regeneration. Each multiple shoot-clump was capable of regenerating more than 80 shoots via an intensive differentiation of both axillary and adventitious shoot buds, the somatic embryos were capable of 90% germination, plant conversion and regeneration. The regenerated shoots could be efficiently rooted on MS medium containing indole-3-butyric acid (IBA 1 mg/l). The plants were successfully transplanted to glasshouse and grown to maturity with a survival rate of 98%. Morphogenetic response of the explants was found to be genotypically independent.     

白花蛇舌草叶片离体培养及试管无性系的建立

以白花蛇舌草叶片为材料,建立了白花蛇舌草叶片高效不定芽发生、植株再生体系。研究了不同激素及其组合对外植体不定芽发生的效应。结果显示,在MS基本培养基中单纯添加6BA,当6BA浓度为0.1mg/L和0.5mg/L时,不能诱导离体叶片发生不定芽,6BA浓度在1.0～5.0mg/L范围内,诱导率随BA浓度升高而增加,适宜浓度为3.0mg/L;当6BA与NAA配合使用时,其诱导率随着培养基中6BA与NAA的相对比值的提高而提高;以MS+BA3mg/L+NAA0.01mg/L作为继代培养基,建立起白花蛇舌草高效、稳定的试管无性系,为白花蛇舌草遗传转化研究奠定了基础。